洗液对拟南芥叶原生质体分离的影响  

The Influence of Washing Buffer on Isolation of Arabidopsis thaliana Mesophyll Protoplasts

作　　者：赵严伟[1] 黄志刚[1] 李合松[1] 

机构地区：[1]湖南农业大学生物科学技术学院,长沙410128

出　　处：《中国农学通报》2011年第12期187-190,共4页Chinese Agricultural Science Bulletin

基　　金：湖南省教育厅科学研究项目(09K051)

摘　　要：原生质体分离过程中洗液对原生质体的纯化和保存起着重要的作用。本试验通过比较30h内WI、W5、MMg3种洗液中原生质体数量与活力的变化以及分析不同浓度CaCl2处理WI、W5、MMg后对原生质体的影响,选择出获得最多活性原生质体数量的洗液。结果表明,WI在维持质膜稳定性上较有优势,而W5则更有利于保持原生质体活力且30h后得到的活性原生质体最多,较适宜在短期内保存原生质体;CaCl2处理洗液对原生质体活力无显著影响,且其对原生质体数量的影响取决于CaCl2处理浓度及洗液种类,MMg洗液在补充0～50mM CaCl2后原生质体数量呈上升趋势。Washing buffer plays an important role in protoplasts purification and storing when protoplasts isolated.The study compared yiled and viability change of protoplasts in washing buffer of WI,W5 and MMg in 30 h,analyzed the influence of CaCl2 with different concentration on protoplasts in WI,W5 and MMg to get the suitable washing buffer for the highest viable protoplasts.The results indicated that WI was better at maintaining plasma membrane,while W5 was better on keeping viability,after 30 h W5 got the highest viable protoplasts population which was good for storing protoplasts in a short period;CaCl2 treatment showed little influence on protoplasts viability and its population improving effect depended on its concentration and the washing buffer type,there was a uptrend of viable protoplasts in MMg when treated with 0-50 mM CaCl2.

关 键 词：原生质体 洗液 CACL2 

分 类 号：Q25[生物学—细胞生物学]